Arble Jessica R, Lalley Andrea L, Dyment Nathaniel A, Joshi Pujan, Shin Dong-Guk, Gooch Cynthia, Grawe Brian, Rowe David, Shearn Jason T
a Biomedical Engineering Program, College of Engineering and Applied Science , University of Cincinnati , Cincinnati , OH , USA.
b Engineered Skin Laboratories , Shriners Hospital for Children , Cincinnati , OH , USA.
Connect Tissue Res. 2016 Nov;57(6):496-506. doi: 10.1080/03008207.2016.1213247. Epub 2016 Aug 23.
Identifying biological success criteria is needed to improve therapies, and one strategy for identifying them is to analyze the RNA transcriptome for successful and unsuccessful models of tendon healing. We have characterized the MRL/MpJ murine strain and found improved mechanical outcomes following a central patellar tendon (PT) injury. In this study, we evaluate the healing of the LG/J murine strain, which comprises 75% of the MRL/MpJ background, to determine if the LG/J also exhibits improved biomechanical properties following injury and to determine differentially expressed transcription factors across the C57BL/6, MRL/MpJ and the LG/J strains during the early stages of healing.
A full-length, central PT defect was created in 16-20 week old MRL/MpJ, LG/J, and C57BL/6 murine strains. Mechanical properties were assessed at 2, 5, and 8 weeks post surgery. Transcriptomic expression was assessed at 3, 7, and 14 days following injury using a novel clustering software program to evaluate differential expression of transcription factors.
Average LG/J structural properties improved to 96.7% and 97.2% of native LG/J PT stiffness and ultimate load by 8 weeks post surgery, respectively. We found the LG/J responded by increasing expression of transcription factors implicated in the inflammatory response and collagen fibril organization.
The LG/J strain returns to normal structural properties by 8 weeks, with steadily increasing properties at each time point. Future work will characterize the cell populations responding to injury and investigate the role of the differentially expressed transcription factors during healing.
为了改进治疗方法,需要确定生物学成功标准,而确定这些标准的一种策略是分析肌腱愈合成功和失败模型的RNA转录组。我们已经对MRL/MpJ小鼠品系进行了特征描述,并发现中央髌腱(PT)损伤后其力学结果有所改善。在本研究中,我们评估了LG/J小鼠品系(其含有75%的MRL/MpJ背景)的愈合情况,以确定LG/J在损伤后是否也表现出改善的生物力学特性,并确定在愈合早期C57BL/6、MRL/MpJ和LG/J品系中差异表达的转录因子。
在16 - 20周龄的MRL/MpJ、LG/J和C57BL/6小鼠品系中制造全长中央PT缺损。在术后2、5和8周评估力学性能。在损伤后3、7和14天使用一种新型聚类软件程序评估转录组表达,以评估转录因子的差异表达。
到术后8周时,LG/J的平均结构性能分别提高到天然LG/J髌腱刚度和极限负荷的96.7%和97.2%。我们发现LG/J通过增加与炎症反应和胶原纤维组织相关的转录因子的表达做出反应。
LG/J品系在8周时恢复到正常结构性能,且在每个时间点性能都在稳步提高。未来的工作将对响应损伤的细胞群体进行特征描述,并研究差异表达的转录因子在愈合过程中的作用。